贵州省PEDV与PoRV分子流行病学调查  

Molecular Epidemiological Investigation of PEDV and PoRV in Guizhou Province

作　　者：丁尊俄 冯旭芳 张双翔[4] 何贤海 胡兴义 张海 王开功[1,2] 文明[1,2] 程振涛 李晨[1,2] 蒲翠敏[1,2] 罗引幸 刘丽娟 周碧君 Ding Zun＇e;Feng Xufang;Zhang Shuangxiang;He Xianhai;Wang Kaigong;Wen Ming;Cheng Zhentao;Li Chen;Pu Cuimin;Luo Yinxing;Liu Lijuan;Zhou Bijun(College of Animal Science, Guizhou University, Guiyang, 550025;Animal Disease Laboratory of Guizhou Province, Guiyang, 550025;Animal Epidemic Disease Control Center of Dejiang in Guizhou Province, Dejiang, 565200;Animal Disease Prevention and Control Center in Guizhou Province, Guiyang, 550001)

机构地区：[1]贵州大学动物科学学院,贵阳550025 [2]贵州省动物疫病研究室,贵阳550025 [3]贵州省德江县动物疫病预防控制中心,德江565200 [4]贵州省动物疫病预防控制中心,贵阳550001

出　　处：《基因组学与应用生物学》2018年第6期2397-2402,共6页Genomics and Applied Biology

基　　金：“贵州省科学技术厅农业公关项目”项目(黔科合NY[2015]3009-1号);“贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队”(黔科合人才团队[2015]4016号)共同资助

摘　　要：为探明贵州省近几年引起猪病毒性腹泻两个主要病原PEDV与PoRV的变异情况,本研究对贵州省贵阳市、遵义市、安顺市、黔东南洲等5个地（州市）26个规模化养猪场采集了213份疑似腹泻的粪便样品,采用实验室建立的PEDV、TGEV和PoRV三重RT-PCR检出PEDV阳性的病料12份和PoRV阳性病料4份。根据GenBank登录的PEDV和PoRV基因序列,分别设计PEDV M基因和PoRV VP7基因特异性引物,扩增目的基因、克隆、测序和遗传进化分析。结果显示：成功克隆了12株PEDV M基因（681 bp）和4株PoRV的VP7基因（981 bp）,M基因核苷酸及推导的氨基酸同源性分别在98.1%～100%和98.2%～100%之间,与国内外参考毒株核苷酸及氨基酸同源性分别在97.4%～100%和97.3%～100%之间;VP7基因核苷酸及推导的氨基酸同源性分别为95.5%～99.7%和96.9%～99.7%,与国内外参考毒株核苷酸及氨基酸同源性分别为71.9%～95.1%和72.7%～99.1%。贵州省PEDV地方流行株与疫苗株CV777、Attenuated DR13亲缘关系较远,PoRV地方流行株与G4型毒株Gottfried、HeN4等属于同一群。结果表明,本研究从分子流行病学角度证实了贵州省PEDV和PoRV流行与变异情况,为贵州省PEDV和PoRV合理的防控措施提供参考依据。In order to find out the variation condition of PEDV and PoRV in two major pathogens of porcine viral diarrhea in recent years, this study was divided Guizhou Province into five regions（states） in Guiyang, Zunyi, Anshun and Qiandongnan, Guizhou province. A total of 213 stool samples of suspected diarrhea were collected from the 26 large-scale pig farms. 12 samples of PEDV positive samples and 4 of PoRV positive samples PEDV and TGEV and PoRV triple RT-PCR were used to detected by using PEDV and TGEV and PoRV triple RT-PCR 12 samples of PEDV positive and 4 of PoRV positive samples. According to the PEDV and PoRV gene sequences registered in GenBank, PEDV M gene and PoRV VP7 gene specific primers were designed to amplify the target gene,cloning, sequencing and genetic evolution analysis. The results showed that the VP7 gene（981 bp） of 12 PEDV M genes（681 bp） and 4 PoRVs were successfully cloned. The nucleotide and deduced amino acid homology of M gene were 98.1%～100% and 98.2%～100%, respectively. The nucleotide and amino acid homology of the reference strains at home and abroad were 97.4%～100% and 97.3%～100%, respectively. The nucleotide and deduced amino acid homology of VP7 gene were 95.5% ～100% and 97.3% ～100%, respectively, and t. he nucleotide and amino acid homology of VP7 gene were 95.5%～99.7% and 96.9%～99.7%, respectively. The nucleotide and amino acid homology of the reference strains were 71.9%～95.1% and 72.7%～99.1%, respectively. The local epidemic strains of PEDV in Guizhou Province were distant from the vaccine strain CV777 and Attenuated DR13, and the local epidemic strains of PoRV belonged to the same group as G4 strains Gottfried and HeN4. The results showed that this study has confirmed the prevalence and variation of PEDV and PoRV in Guizhou Province from the point of molecular epidemiology, and provided the reference for the reasonable prevention and control measures of PEDV and PoRV in Guizhou Province.

关 键 词：PEDV PoRV 基因序列 亲缘关系 

分 类 号：S852.65[农业科学—基础兽医学]